LOCUS       sequence-462850-        5642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human ZDHHC19 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5642)
  AUTHORS   Zhang Z, Ren C, Xiao R, Ma S, Liu H, Dou Y, Fan Y, Wang S, Zhan P, 
            Gao C, Yue X, Li C, Gao L, Liang X, Wu Z, Ma C
  TITLE     Palmitoylation of TIM-3 promotes immune exhaustion and restrains 
            antitumor immunity.
  JOURNAL   Sci Immunol. 2024 Nov 15;9(101):eadp7302. doi: 
            10.1126/sciimmunol.adp7302. Epub 2024 Nov 15.
  PUBMED    39546589
REFERENCE   2  (bases 1 to 5642)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci 
            Immunol."; date: "2024-11-15"; volume: "9(101):eadp7302. doi"; 
            pages: " 10.1126/sciimmunol.adp7302. Epub 2024 Nov 15"
COMMENT     Sequence Label: Flag-ZDHHC19
FEATURES             Location/Qualifiers
     source          1..5642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1983..2605
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2182..2201)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2629..2649)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2634..2963
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2814..2949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3001..3020)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3002..3020)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3034..3050)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3034..3050)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3047..3069)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3058..3074
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3082..3088,3089..3106,3107..3112))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3127..3148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3265..3282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3436..4024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3516..3535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4195..4986,4987..5055))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4818..4837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5056..5160)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5187..5615
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5274..5293)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5484..5505
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagcttg cggccgcgaa ttcaatgaca
     1021 ctcttaacgg atgccacgcc gctggtgaag gagccccatc ccctgcctct ggtcccacgt
     1081 ccctggttcc tccctagcct ctttgctgcc ttcaatgtgg tgctgctggt ctttttcagt
     1141 ggcctcttct tcgcattccc ttgcaggtgg ctggctcaga acggggagtg ggcctttcct
     1201 gttatcacag gctccctctt tgtccttacc ttcttcagtc ttgtttcact caacttctca
     1261 gaccctggca tcttacatca aggctccgct gagcagggcc ccttgacggt gcacgtggtg
     1321 tgggtgaacc acggggcctt ccgcctgcaa tggtgtccaa agtgctgctt ccaccgcccg
     1381 ccccggactt accactgccc ctggtgcaac atctgtgtgg aggactttga ccaccactgc
     1441 aagtgggtca ataactgcat cggtcaccgc aacttccgct tcttcatgct gcttgtcctg
     1501 tccctgtgcc tctactcggg cgccatgctg gtcacctgtc tcatcttcct ggtgcgcaca
     1561 acccacctgc ccttctccac cgacaaggcc atcgccatcg tggtggccgt gtccgccgcg
     1621 ggcctcctgg tgccgctgtc cctcctgctg ctgatccagg cactgtccgt gagctcggcc
     1681 gaccgcacct acaagggcaa gtgcagacac cttcagggat acaacccctt cgaccagggc
     1741 tgtgccagca actggtattt aacaatttgt gcaccactgg gacccaagta catggctgaa
     1801 gctgtccagc tgcagagagt ggtggggcct gactggacat ccatgccgaa tctgcaccct
     1861 ccaatgtccc cctctgctct caacccccca gccccaacct ctgggtccct acaaagcagg
     1921 gaagggaccc ccggggcgtg gtgaagatct gatatcggta ccagtcgact ctagaggatc
     1981 ccgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     2041 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     2101 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     2161 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     2221 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     2281 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     2341 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     2401 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     2461 tgattttaaa ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc
     2521 atgcccaacc ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt
     2581 ccactcagta gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc
     2641 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2701 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2761 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2821 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2881 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2941 ttggaggcct aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt
     3001 ccctatagtg agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     3061 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3841 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5161 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5221 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5281 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5341 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5401 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5461 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5521 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5581 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5641 tt
//
